Incorporation of [14C]glucosamine and [14C]leucine into mouse kidney β-glucuronidase induced by gonadotrophin

Male BALB/C mice were injected intraperitoneally with 2.5 i.u. of gonadotrophin. After the injection, increase of β-glucuronidase activity was first observed in the microsomal fraction. By 36h 45–50% of the total homogenate activity was found in the microsomal fraction compared with 20–25% in the control microsomal fraction. From 36 to 80h not only microsomal β-glucuronidase but also lysosomal β-glucuronidase increased progressively. After 69h stimulation with 2.5 i.u. of gonadotrophin, d-[1-¹⁴C]glucosamine or l-[U-¹⁴C]leucine was injected intraperitoneally. After a further 3h the kidneys were homogenized and five particulate fractions were prepared by differential centrifugation. The β-glucuronidase in the microsomal and lysosomal fractions was released respectively by ultrasonication and by freezing and thawing treatment. The enzyme was purified by organic-solvent precipitation and by sucrose-density-gradient centrifugation. The results demonstrated the incorporation of these two labels into the mouse renal β-glucuronidase. The microsomal β-glucuronidase was much more radioactive than the lysosomal enzyme and approx. 80% of the newly synthesized enzyme appeared in microsomes and approx. 20% of that was found in lysosomes at this period. These results suggest that the mouse renal β-glucuronidase is a glycoprotein and that the newly synthesized enzyme is transported from endoplasmic reticulum to lysosomes.     

RT-PCR检测HUTP14A mRNA时排除假基因HUTP14C干扰的方法

人类U3蛋白14C基因(HUTP14C)是人类U3蛋白14A基因(HUTP14A)的假基因。两者转录本序列同源性高达95%。常规RT-qPCR技术在检测HUTP14A mRNA丰度时,HUTP14C的存在会影响检测结果。本研究旨在建立检测HUTP14A mRNA时排除HUTP14C干扰的RT PCR方法。本研究设计出能分别从多种肿瘤细胞DNA和RNA中特异性扩增HUTP14A和HUTP14C的引物,避免假基因HUTP14C对其同源基因HUTP14A检测的干扰。在检测细胞系HUTP14A mRNA时,通过DNaseⅠ消除RNA中污染的HUTP14C DNA,用靶向HUTP14C 3′-UTR的siRNA沉默HUTP14C mRNA后,再用RT PCR检测HUTP14A mRNA丰度,使结果更加准确。在18对肝癌及癌旁组织中,利用特异性引物进行RT PCR检测,HUTP14A和HUTP14C mRNA的表达略高于癌旁组织。本研究提示,针对有假基因存在的功能基因,对其mRNA丰度进行检测时,在提取细胞或组织总RNA后,用DNaseⅠ处理,再用RNA直接进行PCR扩增,排除DNA污染后,再进行RT-PCR或RT-qPCR扩增。大多假基因具有较长的3′-UTR区,在该区域设计siRNA特异性沉默假基因的mRNA后,用RT-qPCR检测功能基因的mRNA丰度,可以排除假基因mRNA的影响。在病理组织中检测功能基因的mRNA丰度时,可以根据假基因和其功能基因的序列差异设计出特异扩增功能基因的引物,从假基因的3′-UTR区设计特异扩增假基因的引物,通过RT-qPCR技术分别检测二者的mRNA。     

芽胞杆菌C14碱性纤维素酶性质的研究

从碱性污泥中分离到一株产碱性纤维素酶活力较高的耐碱性芽胞杆菌,所产的酶为单一的羧甲基纤维素酶,其专一性底物为羧甲基纤维素钠,酶的最适反应ＰＨ为９．０,在ＰＨ７－１０．０范围内较稳定,酶作用的最适温度为５５℃,在５０℃时较稳定,在６０℃处理,１０、３０、６０和９０分钟后,残余酶活分别为９１．５％、７２．０％、３６．６％和１０．３％。Ｃｏ＾２＋,ＥＤＴＡ对酶促反应有明显的促进作用,Ｈｇ＾２＋,Ａｇ＾２     

14C检测发现了人类心肌细胞更新的证据

长久以来人们一直认为,人类的心脏和大脑一样,在形成后就不再产生新的细胞.然而Bhardwaj等通过对心脏细胞进行14C测定,证实了人类的心肌细胞可以自我更新.     

Assay of Δ1-piperideine-2-carboxylate and synthesis of l-[14C]pipecolate from dl-[14C]pipecolate

Four assay methods were tested for the measurement of Δ¹-piperideine-2-carboxylate, a proposed alicyclic ketimino acid intermediate in the pathway of lysine metabolism to l-pipecolate, and the product of d-amino acid oxidase on d-pipecolate. The method using Δ¹-piperideine-2-carboxylate reductase from Pseudomonas putida was found to be most sensitive and specific. Measurement of Δ¹-piperideine-2-carboxylate by reduction with NaBH₄ and ninhydrin assay of the resultant pipecolate, by direct acidic ninhydrin assay, and by o-aminobenz-aldehyde assay were less desirable because of lower sensitivity and specificity. Two synthetic methods for preparing l-[¹⁴C]pipecolate from the racemic dl-[¹⁴C]pipecolate were investigated. Incubation of dl-[¹⁴C]pipecolate with a combination of d-amino acid oxidase and Δ¹-piperideine-2-carboxylate reductase or d-amino acid oxidase and NaBH₄ totally inverted the d-isomer to the l-isomer, with $\text{Δ}{}^1\text{-[}{}^{14}\text{C]piperideine-2-carboxylate}$ as an intermediate in each cycle of interconversion. No purification except desalting through a Dowex 50 (H⁺) column was necessary in order to recover l-[¹⁴C]pipecolate in pure form. The yield was 95–97% compared to <50% in the conventional method.     

14C示踪技术在土壤有机质周转研究中的应用

采用碳同位素标记有机材料能够较真实地反映其在土壤中的分解和转化过程,是研究土壤有机质周转动力学的必要方法。本文介绍了^14C示踪技术在土壤腐殖质的形成与分解过程、有机底物在土壤中的分解和转化及其对原有土壤有机质分解的影响、土壤微生物生物量碳及其周转以及温室气体排放等方面研究中的应用进展。     

A procedure for the synthesis of p-fluorosulfonyl[14C]-benzoyl-5′-adenosine with [14C] in the benzoyl moiety

Carboxy-p-fluorosulfonyl[¹⁴C]benzoyl-5′-adenosine has been synthesized with the radiolabel ultimately derived from carboxy-p-amino[¹⁴C]benzoic acid by a synthetic route employing four reaction steps. Starting with 1 mmol of p-amino[¹⁴C]benzoic acid, p-fluorosulfonyl[¹⁴C]benzoyl-5′-adenosine is obtained with an overall yield of 25–30%.     

中国真猛犸象和披毛犀化石14C年代研究新进展

真猛犸象（Mammuthus primigenius）和披毛犀（Coelodonta antiquitatis）是北半球高纬度地区晚更新世动物群的主要成员,其消亡的年代和原因一直是国际学术界关注的热点科学问题。本文对黑龙江青冈县英贤村最新出土的5个真猛犸象和5个披毛犀化石进行了AMS¹⁴C年代测定,结果均大于4万年,部分化石可能已经超出了目前¹⁴C的测定范围。通过整理并对比已公开发表的中国境内两种动物化石的¹⁴C年代学数据,本文认为早期常规¹⁴C测年方法所获得的年代值需要重新考虑其准确性。埋藏地层与最新的AMS¹⁴C测年数据显示,我国真猛犸象化石年代主要集中于MIS3阶段;披毛犀在我国消亡的时间很可能晚于真猛犸象,至少延续到末次冰消期。中国猛犸象-披毛犀动物群化石仍然需要开展更多的年代学研究。     

测定浮游植物光合作用速率的~(14)C方法

如何提高测定浮游植物光合作用速率的准确性,一直是一个引起讨论的问题。放射性同位素法(~(14)C法)虽然被认为是比较好的方法,但其在国内尚未得到广泛地应用。我们在实验中发现,用一般直接制样技术的     

地层的年龄和~(14)C年代测定法

在中学生物学课本中,有关生物进化的章节都附有一个“地质年代表”。在“距今年数”栏目里开列出了各地质时代(代、纪、世等)距离现代的年数。如中生代从距今2亿2千5百万年开始等等。有的学生向教师提出,这些年代是如何计算出来的?回答这个问题,还得从放射性同位素谈起。因为表里所列出的年代是地层形成的绝对年龄。它是根据放射性元素衰变规律计算出来的。     

Incorporation of label from d-β-hydroxy-[14C]butyrate and [3-14C]acetoacetate into amino acids in rat brain in vivo

The metabolism of ketone bodies by rat brain was studied in vivo. Rats starved for 48h were given either d-beta-hydroxy[3-(14)C]butyrate or [3-(14)C]acetoacetate by intravenous injection and killed after 3 or 10min. Total radioactivity in the acid-soluble material of the brain and the specific radioactivities of the brain amino acids glutamate, glutamine, aspartate and gamma-aminobutyrate were determined. A group of fed animals were also given d-beta-hydroxy[3-(14)C]butyrate. In the brains of all animals (14)C was present in the acid-soluble material and the specific radioactivity of glutamate was greater than that of glutamine.     

A C14-polyacetylenic glucoside with an α-pyrone moiety and four C10-polyacetylenic glucosides from Mediasia macrophylla

Polyacetylenic glucosides (1–5) were isolated from the MeOH extract of Mediasia macrophylla, and their structures were established by spectroscopic analyses. Compounds 2–4 were the first examples of C₁₀-polyacetylenic glucosides found in the family Umbelliferae, while compound 1 was a unique polyacetylenic glucoside possessing an α-pyrone moiety.     

雷公藤内酯醇C14位羟基结构修饰及抗肿瘤活性研究进展

雷公藤是我国资源丰富的一种传统中草药,具有抗炎、抗肿瘤、免疫抑制等多种生物活性。本文综述了近年来雷公藤内酯醇C14位羟基的结构修饰和抗肿瘤活性的研究进展。     

生物样品中~(14)C 的氧瓶燃烧测定法

制备同位素生物样品进行液体闪烁测定的方法很多。氧瓶燃烧法由于操作简便,样品取量大,且最终得到的是无色液体,因而能克服一些比放射性低样品在测定时的化学焠灭;或在消化处理样品时引起~(14)CO_2的丢失等缺点,目前愈来愈广泛地得到重视和应用。我们在进行~(14)C-棉酚的吸收、分布和排泄研究时,曾采用过     

Sclerosponge growth rate as determined by 210Pb and Δ14C chronologies

Measurements of bomb-produced radiocarbon and ²¹⁰Pb provide concordant estimates of the growth rate of the sclerosponge Ceratoporella nicholsoni collected from the reef slope of northern Jamaica. Radiocarbon measurements of older growth bands in the same specimen are similar to the time history of radiocarbon in coral bands from two sites in the northwestern Atlantic. Furthermore, ²¹⁰Pb and stable Pb analyses reveal that the sclerosponge incorporates this element at much higher concentrations than corals.Contribution No. 5955 to the Woods Hole Oceanographic Institution     

~（14）C－涕灭威在旱田土壤中的降解

研究了１４Ｃ－涕灭威在５种土壤中（４ｐｐｍ,互．２２μＣｉ·５０ｇ－１土壤干重）的生物降解．模拟试验为密闭系统,土壤中水分含量为２２％,气温２０—３０Ｃ℃在供试的５种土壤中,北京肖家河的土壤降解最快,为施人放射剂量的５１．３％,以“ＣＯ：形式从土壤进出;２６．０％与土壤结合,只有２１．６％可以被抽出．取自浙江义乌的土壤降解较慢,收集到的１４ＣＯ２为施入量的２３．３％．土壤中加入杀菌剂红霉素或敌茵丹降解作用明显减慢．土壤提取物中涕灭威亚砜、涕灭威亚砜肟被确认是主要的代谢产物,还发现了少量的涕灭威砜,涕灭威亚砜腈涕灭威砜腈和涕灭威砜肟等降解物．     

Bomb <Superscript>14</Superscript>C enrichment indicates decadal C pool in deep soil?

Studies of changes in soil organic carbon (SOC) stocks normally limit their focus to the upper 20–30 cm of soil, yet 0–20 cm SOC stocks are only ∼40% of 0–1 m SOC. Accounting for only the upper 20–30 cm of SOC has been justifiable assuming that deeper SOC is unreactive since it displays ¹⁴C-derived mean residence times of hundreds or thousands of years. The dramatic increase in the ¹⁴C content of the atmosphere resulting from thermonuclear testing circa 1963 allows the unreactivity of deep SOC to be tested by examining whether deep soils show evidence of ‘bomb-¹⁴C’ incorporation. At depths of 40–100 cm, a well-studied New Zealand soil under stable pastoral management displays progressive enrichment of over 200‰ across samplings in 1959, 1974 and 2002, indicating substantial incorporation of bomb ¹⁴C. This pattern of deep ¹⁴C enrichment—previously observed in 2 well-drained California grassland soils—leads to the hypothesis that roots and/or dissolved organic C transport contribute to a decadally-reactive SOC pool comprising ∼10–40% of SOC below 50 cm. Deep reactive SOC may be important in the global C cycle because it can react to land-use or vegetation change and may respond to different processes than the reactive SOC in the upper 20–30 cm of soil.